Inverse PCR (or alternative) recommendations requested

Paul N Hengen pnh at cockleberry.ncifcrf.gov
Mon Dec 4 13:15:10 EST 1995


Zunde Wang (U16788 at uicvm.uic.edu) wrote:

> I am considering using inverse PCR to attempt to locate transgenes
> in the mouse genome.

: I am now considering using linker ligations. PanVera sells a PCR
: cloning kit which has six sticky-ended linkers and can be ligated to
: DNA digested with seven restriction enzymes. Since it's quite expensive
: ($350 for 20 reactions), I'd like to ask those who have used it how
: effective the kit is in flanking sequence cloning.

Uhhh...why by a kit for linkers when you can make your own for less?
I suppose it would save you the thought process, but you can do this
without using a kit. Have you considered using vectorettes and a primer
specific to the gene of interest? You only need sequence at one end, right?

@article{Hengen1995Septibs,
author = "P. N. Hengen",
title = "Methods and reagents - Vectorette, splinkerette,
and boomerang {DNA} amplification",
journal = "Trends in Biochemical Sciences",
volume = "20",
number = "9",
pages = "372-373",
month = "sep",
year = "1995"}

*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - - - - - Methods FAQ list -> http://www-lmmb.ncifcrf.gov/~pnh/ - - - - - - *
* - - -  Anonymous FTP from ftp.ncifcrf.gov as file pub/methods/FAQlist - - - *
*******************************************************************************



More information about the Methods mailing list